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realtime from different amounts of starting tissue mass

realtime from different amounts of starting tissue mass - Real-Time PCR and Quantitative PCR Forum

realtime from different amounts of starting tissue mass - Real-Time PCR and Quantitative PCR Forum


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Old 11-22-2007, 07:58 AM
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Question realtime from different amounts of starting tissue mass



Hi all,

i would like to do some real time pcr over the life course of an insect
I would like to measure gene expression in the larvae, pupae, adult body and adult antennae.

i would probably use whole larvae, pupae and adult bodies, but the adult antennae are much much smaller amounts of tissue than the others

is it fair to do realtime on these tissues and compare gene expression across them, even though one tissue sample has a much smaller mass than the others?
of course i would be adding the same amount of extracted RNA to the cDNA synthesis, but being from different tissue masses, would i expect the cts of the housekeeping genes to vary? (some more dilute than others?) could you use houskeeping cts to normalise againsnt and would this normalisation correct for the difference in starting tissue mass?

i'm abit confused with it all so hope someone can run me through it!!

thanks alot!!
GF820
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